高糖对人晶状体上皮细胞硫醇转移酶表达的影响

发布时间：2018-07-31 08:22

【摘要】：目的观察高浓度葡萄糖对人晶状体上皮细胞硫醇转移酶(thioltransferase,TTase)表达的影响。方法将体外培养的人晶状体上皮细胞分别用葡萄糖浓度为5.5 mmol·L-1(正常对照组)、15.5 mmol·L-1、25.5 mmol·L-1及35.5 mmol·L-1(不同浓度高糖组),胎牛血清体积分数均为15%的DMEM培养液培养,同时设5.5 mmol·L-1葡萄糖+30.0 mmol·L-1甘露醇高渗对照组,24 h后Western-blotting检测TTase表达情况。选择35.5 mmol·L-1高糖浓度培养细胞,分别在0 h、2 h、8 h、16h、24 h时间点收集细胞,Western-blotting检测TTase表达,并测定TTase活性,细胞内总谷胱甘肽(T-GSH)、还原型谷胱甘肽(GSH)与氧化型谷胱甘肽(GSSG)含量,计算GSSG与T-GSH的比值。结果 TTase表达随葡萄糖浓度增加逐渐增强,在35.5mmol·L-1高糖组中表达最强,约为正常对照组的2.3倍(P0.01),25.5 mmol·L-1高糖组约为正常对照组1.7倍(P0.05),甘露醇高渗对照组TTase表达与正常对照组相比无明显变化(P0.05)。在35.5 mmol·L-1高糖处理后2 h,TTase表达开始增高,8 h表达达到高峰,约为正常对照组的2.0倍(P0.01),随后逐渐下降,16 h时约为正常对照组1.7倍(P0.01);24 h降为1.5倍(P0.05)。与TTase表达类似,TTase活性在35.5 mmol·L-1高糖处理后2 h开始升高,8 h达到高峰,约为正常对照组的1.8倍(P0.01),随后逐渐下降。GSH在35.5 mmol·L-1高糖处理后16 h较对照组明显下降(P0.05)。GSSG和GSSG/T-GSH比值在35.5 mmol·L-1高糖处理2 h后均显著升高,分别约为正常对照组的3.5倍(P0.001)和3.3倍(P0.001),并随时间推移不断上升。结论高糖可诱导晶状体上皮细胞TTase表达上调、活性增强,同时伴GSH的含量下降,GSSG含量上升。
[Abstract]:Objective to observe the effect of high concentration glucose on the expression of thioltransferase (TTase) in human lens epithelial cells. Methods cultured human lens epithelial cells in vitro were cultured in DMEM medium with glucose concentration of 5.5 mmol L-1 (normal control group), 15.5 mmol L -1 and 35.5 mmol L -1 (different concentrations of high glucose), respectively, and the volume fraction of fetal bovine serum was 15%. At the same time, 5.5 mmol L-1 glucose 30.0 mmol L-1 mannitol hypertonic control group 24 h after Western-blotting detection of TTase expression. The cells cultured at high glucose concentration (35.5 mmol L-1) were collected at 0 h, 8 h and 16 h for 24 h. The expression of TTase was detected by Western-blotting. The activity of TTase, the content of total glutathione (T-GSH), reduced glutathione (GSH) and oxidative glutathione (GSSG) were measured. The ratio of GSSG to T-GSH was calculated. Results the expression of TTase increased gradually with the increase of glucose concentration, and the expression of TTase was the strongest in 35.5mmol L-1 high glucose group. It was about 2.3-fold (P0.01) of normal control group (P0.01) and about 1.7-fold of normal control group in 25.5 mmol L-1 high glucose group (P0.05). The TTase expression of mannitol hyperosmotic control group had no significant change compared with normal control group (P0.05). After 35.5 mmol L ~ (-1) high glucose treatment, the expression of TTase began to increase to a peak at 8 h, which was about 2.0-fold (P0.01) of the normal control group, and then decreased to 1.5-fold (P0.01) at 16 h after the treatment compared with that of the normal control group (P0.01) at 24 h. After 35.5 mmol L ~ (-1) high glucose treatment, the activity of TTase was increased to a peak at 8 h after treatment with 35.5 mmol L ~ (-1) high glucose. Compared with the control group, the ratio of GSSG and GSSG/T-GSH increased significantly after 35.5 mmol L ~ (-1) high glucose treatment for 2 h (P0.05), and the ratio of GSSG to GSSG/T-GSH was significantly increased after 35.5 mmol L ~ (-1) high glucose treatment for 2 h after treatment with 35.5 mmol 路L ~ (-1) high glucose (P _ (0.01). It was about 3.5 times (P0.001) and 3.3 times (P0.001) as compared with the normal control group, and increased with time. Conclusion High glucose can induce the up-regulation of TTase expression and increase of activity in lens epithelial cells, while the content of GSH decreases and the content of GSSG increases.
【作者单位】：第四军医大学唐都医院眼科;
【基金】：国家自然科学基金资助(编号:81070720)~~
【分类号】：R587.2;R776.1

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